FUNCTIONAL EXPRESSION OF A VERTEBRATE INWARDLY RECTIFYING K+ CHANNEL IN YEAST
FUNCTIONAL EXPRESSION OF A VERTEBRATE INWARDLY RECTIFYING K+ CHANNEL IN YEAST
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DOI：
10.1091/mbc.6.9.1231
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发表时间：
1995-09-01
影响因子：
3.3
通讯作者：
KURTZ, S
中科院分区：
文献类型：
--
作者：
TANG, WM;RUKNUDIN, A;KURTZ, S
我们描述了gpIRK 1的表达，gpIRK 1是从豚鼠心脏cDNA中获得的一种内向整流K+通道。gpIRK 1是在巨噬细胞中鉴定的小鼠IRK 1通道的同源物。gpIRK 1在非洲爪蟾卵母细胞中的表达产生内向整流K+电流，类似于心脏内向整流电流I-K1。该电流被外部Ba 2+和Cs+阻断。构建了在组成型（PGK）或诱导型（GAL）启动子的转录控制下含有gpIRK 1编码区的质粒，用于在酿酒酵母中表达。一些观察结果表明，gpIRK 1在酵母中表达时形成功能性离子通道。gpIRK 1补充了trk 1 Δ trk 2 Δ菌株，该菌株在钾摄取方面有缺陷。gpIRK 1在该突变体中的表达恢复了在低钾培养基上的生长。依赖于gpIRK 1的生长被外部Cst抑制表达gpIRK 1的菌株为研究内向整流Kt通道的组装和组成提供了一个通用的遗传系统。
We describe the expression of gpIRK1, an inwardly rectifying K+ channel obtained from guinea pig cardiac cDNA. gpIRK1 is a homologue of the mouse IRK1 channel identified in macrophage cells. Expression of gpIRK1 in Xenopus oocytes produces inwardly rectifying K+ current, similar to the cardiac inward rectifier current I-K1. This current is blocked by external Ba2+ and Cs+. Plasmids containing the gpIRK1 coding region under the transcriptional control of constitutive (PGK) or inducible (GAL) promoters were constructed for expression in Saccharomyces cerevisiae. Several observations suggest that gpIRK1 forms functional ion channels when expressed in yeast. gpIRK1 complements a trk1 Delta trk2 Delta strain, which is defective in potassium uptake. Expression of gpIRK1 in this mutant restores growth on low potassium media. Growth dependent on gpIRK1 is inhibited by external Cst The strain expressing gpIRK1 provides a versatile genetic system for studying the assembly and composition of inwardly rectifying Kt channels.